Modified vitamin K-dependent polypeptides

Abstract

The invention provides vitamin K-dependent polypeptides with enhanced membrane binding affinity. These polypeptides can be used to modulate clot formation in mammals. Methods of modulating clot formation in mammals are also described.

What is claimed is: 
     
       1. A method of decreasing blood clot formation, wherein said method comprises administering to a patient in need thereof a protein C or activated protein C polypeptide comprising a modified gamma-carboxylglutamic acid (GLA) domain, said modified GLA domain comprising the amino acid sequence of SEQ ID NO:1 with two, three, four, or five amino acid substitutions at positions selected from the group consisting of residues 10, 11, 28, 32, and 33, thereby decreasing blood clot formation in said patient. 
     
     
       2. The method of  claim 1 , wherein said modified GLA domain comprises the amino acid sequence of SEQ ID NO:1 with three amino acid substitutions at residues 11, 32, and 33. 
     
     
       3. The method of  claim 2 , wherein residue 32 of SEQ ID NO:1 is glutamic acid and residue 33 of SEQ ID NO:1 is aspartic acid. 
     
     
       4. The method of  claim 2 , wherein residue 11 of SEQ ID NO:1 is glycine. 
     
     
       5. The method of  claim 1 , wherein said modified GLA domain comprises the amino acid sequence of SEQ ID NO:1 with three amino acid substitutions at residues 11, 32, and 33, wherein residue 11 of SEQ ID NO: 1 is glycine, residue 32 of SEQ ID NO:1 is glutamic acid, and residue 33 of SEQ ID NO:1 is aspartic acid. 
     
     
       6. The method of  claim 1 , wherein said method comprises administering said activated protein C polypeptide.